Nitrogen-containing biphenyl compounds, pharmaceutical compositions of same, preparation methods and anti-HIV-1 uses thereof

ABSTRACT

Nitrogen-containing biphenyl compounds as represented by formula (I), pharmaceutically acceptable salts or derivatives thereof, pharmaceutical compositions, and preparation methods therefore, and anti-HIV-1 use of the compound. Each substituent group in formula (I) is as defined in the description.

This U.S. patent application claims the benefit of PCT application no.PCT/CN2011/080718, filed on Oct. 13, 2011, which claims priority toChinese Application Ser. No. 201010512452, filed Oct. 20, 2010. Theentire contents of the aforementioned applications are incorporatedherein by reference in their entireties.

TECHNICAL FIELD

The present invention belongs to pharmaceutical field and in particularrelates to nitrogen-containing biphenyl compounds having the structuralformulae (I) and (II), pharmaceutical compositions comprising thecompounds as active ingredient and a pharmaceutically acceptablecarrier, methods for their preparation and their use in the preparationof anti-HIV (HIV-1) drug.

BACKGROUND ART

Since the first case report in 1981, AIDS quickly spreads to countriesaround the world. So far, there is still no available effective anti-HIVvaccine, so the development of anti-AIDS drugs is still the mostprospective route for the treatment of AIDS. At present, anti-AIDS drugsmainly include the following three categories: nucleotide antiviraldrugs, transcriptase inhibitors and protease inhibitors. However,because of their high price (more than two or three hundred thousandYuan is needed for each person each year), and relatively great sideeffects, these drugs are beyond the reach of the average family.Therefore, it is urgent to develop an anti-AIDS drug that is simple touse, low price, highly potent, and low toxicity. In recent years,natural products have shown promising results in the treatment of AIDS.Until now, it has been that there are more than 100 kinds of naturalproducts which have good anti-HIV activity, such as: glycyrrhizin,hypericin, curcumin, soy saponins, camptothecin, lentinan, smallpoxprotein and so on.

Lignans, which are isolated from a traditional Chinesemedicine—Schisandra by the applicant in recent years, show a very stronginhibitory effect on HIV integrase, and are able to inhibit thetranscription of HIV-1, which can be developed to be anti-AIDS drugs(CN1634031A; CN1931151A; CN101318950A). Also, these natural products canbe used as a lead compound, and serve as a template for the design ofnew drugs having a stronger biological activity. To date, there is noreport in the prior art relating to nitrogen-containing biphenylcompounds, nor is there any report relating to use of this class ofcompounds as anti-AIDS drugs.

CONTENTS OF THE INVENTION

The object of the present invention is to provide a class ofnitrogen-containing biphenyl compound having anti-HIV activity, toprovide a pharmaceutical composition comprising said compound and apharmaceutically acceptable carrier, and to provide methods forpreparing said compound and its pharmaceutical composition. Anotherobject of the present invention is to provide use of thenitrogen-containing biphenyl compound in the preparation of a drug forthe treatment and prevention of AIDS. Still another object of thepresent invention is to provide a method for the treatment andprevention of AIDS, which method comprises administering to patients thenitrogen-containing biphenyl compound of the present invention.

The above objects of the present invention are realized by the followingtechnical solutions:

The present invention provides a nitrogen-containing biphenyl compoundhaving the following structural formula (I) or (II):

wherein

R₁, R₂, R₃ and R₄ are selected from the group consisting of H, OH,OCH₂Ph, OR, F, Cl, Br and I, or R₁ and R₂ together form —OCH₂O—, or R₂and R₃ together form —OCH₂O—, or R₃ and R₄ together form —OCH₂O—;

R₅ is selected from the group consisting of H, aliphatic hydrocarbyl,aromatic hydrocarbyl, OH, OCH₂Ph, OR and NR₂;

R₆ and R₇ are selected from the group consisting of H, aliphatichydrocarbyl containing 1-15 carbon atoms, CH₂Ph, aromatic hydrocarbyland RCO, or R₆ and R₇ together form —(CH₂)_(n)—, wherein n=2-10, or R₆and R₇ together form —CH₂CH₂OCH₂CH₂—;

R₈ is selected from the group consisting of H, aliphatic hydrocarbyl,CH₂Ph and aromatic hydrocarbyl;

R₉ is selected from the group consisting of H, aliphatic hydrocarbyl andaromatic hydrocarbyl;

R is selected from the group consisting of aliphatic hydrocarbyl andaryl;

X is selected from the group consisting of F, Cl, Br, I, OH, SO₄ andNO₃.

In the nitrogen-containing biphenyl compound, the nitrogen atom linkedto the radicals R₆, R₇ is located in any position of the benzene ringwhere it is present.

In one embodiment of the present invention, R₁ is selected from thegroup consisting of OH, OCH₂Ph and OR, preferably OCH₃ or OCH₂CH₃; R₂and R₃ together form —OCH₂O—; R₅ is selected from the group consistingof OCH₂Ph, OCH₃ and OCH₂CH₃; R₆ and R₇ are selected from the groupconsisting of aliphatic hydrocarbyl containing 1-10 carbon atoms; R₈ isselected from the group consisting of H, CH₂Ph and aliphatichydrocarbyl; R₉ is selected from the group consisting of H and aliphatichydrocarbyl; X is Cl.

In one embodiment of the present invention, R₁ is OCH₃.

In one embodiment of the present invention, R₂ and R₃ together form—OCH₂O—.

In one embodiment of the present invention, R₅ is OCH₃.

In one embodiment of the present invention, R₆ and R₇ are methyl.

In one embodiment of the present invention, R₈ is CH₂Ph.

In one embodiment of the present invention, R₉ is methyl.

In one embodiment of the present invention, X is Cl.

DEFINITIONS

As used herein, the term “aliphatic hydrocarbyl” means a straight-chain(i.e., non-branched) or branched hydrocarbon chain that is completelysaturated or that contains one or more units of unsaturation but isnon-aromatic. Unless otherwise specified, the aliphatic hydrocarbylcontains 1-15 aliphatic carbon atoms (C₁₋₁₅ aliphatic hydrocarbyl). Inone embodiment, the aliphatic hydrocarbyl contains 1-10 aliphatic carbonatoms (C₁₋₁₀ aliphatic hydrocarbyl). In another embodiment, thealiphatic hydrocarbyl contains 1-8 aliphatic carbon atoms (C₁₋₈aliphatic hydrocarbyl). In still another embodiment, the aliphatichydrocarbyl contains 1-6 aliphatic carbon atoms (C₁₋₆ aliphatichydrocarbyl). In still another embodiment, the aliphatic hydrocarbylcontains 1-4 aliphatic carbon atoms (C₁₋₄ aliphatic hydrocarbon). Thealiphatic hydrocarbyl may be straight chain or branched chain alkyl,alkenyl or alkynyl group. Specific examples of aliphatic hydrocarbylinclude, but are not limited to, methyl, ethyl, n-propyl, isopropyl,n-butyl, tert-butyl, vinyl, n-butenyl, ethynyl and the like.

As used herein, the term “alkyl” means a saturated straight-chain orbranched-chain hydrocarbyl. As used herein, the term “alkenyl” refers toa straight-chain or branched-chain hydrocarbyl containing one or moredouble bonds. As used herein, the term “alkynyl” means a straight-chainor branched-chain hydrocarbyl containing one or more triple bonds.

As used herein, the term “aromatic hydrocarbyl” means an unsaturatedaromatic carbocyclic group having one single ring or two or more fusedrings having 6-18 carbon atoms (C₆₋₁₈ aromatic hydrocarbyl). Thearomatic hydrocarbyl preferably has 6-10 carbon atoms (C₆₋₁₀ aromatichydrocarbyl), more preferably having 6 carbon atoms (C₆ aromatichydrocarbyl). Typical examples of the “aromatic hydrocarbyl” include,but are not limited to, phenyl, naphthyl, anthryl and the like.

The preferred nitrogen-containing biphenyl compounds of the presentinvention are compounds 5-12 having the following structural formulae:

The present invention also provides a method for the preparation of thenitrogen-containing biphenyl compound of the structural formula (I),which comprises:

coupling Compound 1 and Compound 2 by metallic catalysis to obtain thenitrogen-containing biphenyl compound of the structural formula (I),wherein the reaction formula is shown as follows:

wherein:

X′ is F, Cl, Br or I;

R′ is an aliphatic or aromatic hydrocarbyl, or two R's together form—OC(CH₃)₂—C(CH₃)₂O—;

the metal catalyst is palladium, platinum, rhodium, ruthenium, lithium,copper, magnesium, nickel or zinc; and

R₁-R₇ are as defined above in the formula (I) and formula (II); and

optionally, the obtained nitrogen-containing biphenyl compound of thestructural formula (I) is subjected to a corresponding salt-formingreaction, to obtain the nitrogen-containing biphenyl compound of thestructural formula (II).

A general illustrative preparation method of the nitrogen-containingbiphenyl compound of the structural formula (I) is described as follows:mixing Compound 1 (0.1 mmol) and Compound 2 (0.2-0.5 mmol), then addingPd(OAc)₂ (0.01-0.05 mmol), Bu₄NBr (0.15 mmol), K₂CO₃ (0.3 mmol) and 3 mLTHF-H₂O (V_(THF)/V_(H2O)=1) to the resultant mixture, heating themixture to about 70° C., reacting for about 4 hours, cooling the mixtureto room temperature, filtering and concentrating the reaction solution,and purifying by a silica gel column, to obtain the compound of thestructural formula (I).

A general illustrative preparation method of the nitrogen-containingbiphenyl compound of the structural formula (II) is described asfollows: the compound of the structural formula (I) obtained by theabove method is subjected to a corresponding salt-forming reaction, toobtain a variety of ammonium salts or quaternary ammonium salts, i.e.,the compounds of the structural formula (II).

The present invention also provides a pharmaceutical composition whichcomprises the compound of the structural formula (I), the compound ofthe structural formula (II), in particular one of the compounds 5-12, ofthe present invention as an active ingredient, and at least onepharmaceutically acceptable carrier.

The present invention also provides use of the nitrogen-containingbiphenyl compound of the present invention in the preparation of a drugfor the treatment or prevention of AIDS.

As used herein, the pharmaceutically acceptable carrier refers to aconventional pharmaceutical carrier in the pharmaceutical field, suchas: diluents, excipients such as water, fillers such as starch, sucroseand the like; binders such as cellulose derivatives, alginates, gelatin,and polyvinyl pyrrolidone; wetting agents such as glycerol;disintegrating agents such as agar, calcium carbonate and sodiumbicarbonate; absorption accelerators such as quaternary ammoniumcompounds; surface active agents such as cetyl alcohol; adsorptioncarriers such as kaolin and bentonite; lubricants such as talc, calciumand magnesium stearate, and polyethylene glycol and the like.Furthermore, other auxiliary agents, such as flavoring agents,sweeteners and the like, can also be added to the composition.

The compound of the present invention can be applied to patients in needof such treatment in the form of composition by oral, nasal inhalation,rectal or parenteral administration. For oral administration, it can bemade into conventional solid preparations such as tablets, powders,granules, capsules and the like, or made into liquid preparations, suchas water or oil suspensions or other liquid preparations such as syrups,elixirs and the like; for parenteral administration, it can be made intoinjection solutions, aqueous or oily suspensions and the like.Preferably, the composition is in the form of tablets, capsules andinjections.

The various dosage forms of the pharmaceutical composition of thepresent invention can be prepared in accordance with conventionalproduction method in pharmaceutical field. For example, the activeingredient is mixed with one or more carriers, and then formed into thedesired dosage form.

The pharmaceutical composition of the present invention comprises theactive ingredient in a weight percent of preferably 0.1%-99.5%, and mostpreferably 0.5%-95%.

The dosage of the compound of the present invention can vary accordingto the route of administration, the patient's age, body weight, the typeand severity of the disease to be treated and the like, and the dailydosage thereof may be 0.01-10 mg/kg body weight, preferably 0.1-5 mg/kgbody weight. It can be administered once or several times.

MODE OF CARRYING OUT THE INVENTION

The following examples are exemplifying, but not limiting, the method ofthe present invention. Other modifications and adaptive changes onvarious conditions and parameters, which are obvious to a person skilledin the art, are all included within the spirit and scope of the presentinvention.

EXAMPLE 1

Preparation method of nitrogen-containing biphenyl compound of thepresent invention can be generally summarized as follows:

Compound 3 (0.1 mmol) and Compound 4 (0.2-0.5 mmol) were put in a 25 mLround-bottomed flask, followed by addition of Pd(OAc)₂ (0.01-0.05 mmol),Bu₄NBr (0.15 mmol), K₂CO₃ (0.3 mmol) and 3 mL THF-H₂O(V_(THF)/V_(H2O)=1). The mixture was heated to 70° C. and the reactionwas carried out for 4 hours. The reaction solution was cooled to roomtemperature, filtered, concentrated, and purified by silica gel columnto give compounds 5-8 in Example 2-5.

Compound 4 is purchased from Acros or Sigma-Aldrich Company.

Compound 3 is obtained by the following synthetic route:

(a) Synthesis of Bromo Compound 3:

Bromo Compound 3 was synthesized using gallic acid as a startingmaterial, through esterification of carboxyl group, etherification ofphenolic hydroxyl group, bromination of benzene ring, removal of benzylprotecting group, and final methyl etherification.

(b) Synthesis of Iodo Compound 3:

Iodo Compound 3 was synthesized using one intermediate compound obtainedduring the synthesis of bromo Compound 3 as a starting material, throughremoval of benzyl protecting group, methyl etherification, nitration ofbenzene ring, reduction of nitro group into amino group, and finalSandmeyer reaction (amino group was firstly converted to a diazoniumsalt in the presence of concentrated HCl+NaNO₂, and then, after theaddition of an aqueous solution of KI, the iodo compound was obtained).

EXAMPLE 2

Bromo Compound 3 (0.1 mmol) obtained in Example 1 and4-(N,N-dimethylamino)phenylboronic acid (CAS: 28611-39-4) (0.2 mmol)were put in a 25 mL round-bottomed flask, followed by addition ofPd(OAc)₂ (0.01 mmol), Bu₄NBr (0.15 mmol), K₂CO₃ (0.3 mmol), and 3 mLTHF-H₂O (V_(THF)/V_(H2O)=1). The mixture was heated to 70° C. and thereaction was carried out for 4 hours. The reaction solution was cooledto room temperature, filtered, concentrated, and purified by silica gelcolumn to give Compound 5.

The relevant test data were as follows: ¹H-NMR (300 MHz, CDCl₃): δ=7.09(d, 2H), 7.03 (s, 1H), 6.74 (d, 2H), 6.02 (s, 2H), 3.77 (s, 3H), 3.57(s, 3H), 2.98 (s, 6H); ¹³C-NMR (75 MHz, CDCl₃): δ=168.40, 149.49,147.67, 141.44, 140.13, 131.16, 130.20, 125.83, 124.16, 111.74, 104.38,101.74, 60.00, 51.90, 40.48.

EXAMPLE 3

Bromo Compound 3 (0.1 mmol) obtained in Example 1 and4-(N,N-dimethylamino)-3-phenylboronic acid (CAS: 919496-59-6) (0.3 mmol)were put in a 25 mL round-bottomed flask, followed by addition ofPd(OAc)₂ (0.02 mmol), Bu₄NBr (0.15 mmol), K₂CO₃ (0.3 mmol), and 3 mLTHF-H₂O (V_(THF)/V_(H2O)=1). The mixture was heated to 70° C. and thereaction was carried out for 4 hours. The reaction solution was cooledto room temperature, filtered, concentrated, and purified by silica gelcolumn to give Compound 6.

The relevant test data were as follows: ¹H-NMR (300 MHz, CDCl₃):δ=6.92-6.95 (m, 4H), 5.97 (s, 2H), 3.70 (s, 3H), 3.45 (s, 3H), 2.66 (s,6H), 2.26 (s, 3H); ¹³C-NMR (75 MHz, CDCl₃): δ=167.92, 149.43, 147.72,145.77, 140.34, 131.68, 127.31, 125.93, 124.65, 123.20, 115.11, 112.24,108.13, 101.54, 56.40, 51.86, 40.55, 16.23.

EXAMPLE 4

Iodo Compound 3 (0.1 mmol) obtained in Example 1 and 4-morpholinylphenylboronic acid pinacol ester (CAS: 568577-88-8) (0.5 mmol) were putin a 25 mL round-bottomed flask, followed by addition of Pd(OAc)₂ (0.05mmol), Bu₄NBr (0.15 mmol), K₂CO₃ (0.3 mmol), and 3 mL THF-H₂O(V_(THF)/V_(H2O)=1). The mixture was heated to 70° C. and the reactionwas carried out for 4 hours. The reaction solution was cooled to roomtemperature, filtered, concentrated, and purified by silica gel columnto give Compound 7.

The relevant test data were as follows: ¹H-NMR (300 MHz, CDCl₃): δ=6.92(s, 1H), 6.03 (s, 2H), 3.85-3.89 (t, 4H), 3.76 (s, 3H), 3.56 (s, 3H),3.19-3.22 (t, 4H); ¹³C-NMR (75 MHz, CDCl₃): δ=168.39, 150.00, 147.92,141.67, 140.78, 128.01, 104.52, 101.83, 66.98, 60.05, 51.95, 49.06.

EXAMPLE 5

Iodo Compound 3 (0.1 mmol) obtained in Example 1 and4-(N,N-diphenylamino)phenylboronic acid (CAS: 201802-67-7) (0.5 mmol)were put in a 25 mL round-bottomed flask, followed by addition ofPd(OAc)₂ (0.05 mmol), Bu₄NBr (0.15 mmol), K₂CO₃ (0.3 mmol), and 3 mLTHF-H₂O (V_(THF)/V_(H2O)=1). The mixture was heated to 70° C. and thereaction was carried out for 4 hours. The reaction solution was cooledto room temperature, filtered, concentrated, and purified by silica gelcolumn to give Compound 8.

The relevant test data were as follows: ¹H-NMR (300 MHz, CDCl₃):δ=6.47-7.32 (m, 15H), 5.98 (s, 2H), 3.76 (s, 3H), 3.62 (s, 3H). ¹³C-NMR(75 MHz, CDCl₃): δ=168.55, 149.50, 148.71, 141.43, 140.32, 139.93,131.43, 130.53, 129.11, 125.76, 124.64, 123.32, 121.11, 113.00, 107.71,101.73, 61.11, 51.87.

EXAMPLE 6

Compound 5 (0.01 mmol) obtained in Example 2 was put in a 10 mLround-bottomed flask, followed by addition of 2 mL methanol and 0.1 mL40% aqueous solution of NaOH. The reaction was carried out at roomtemperature with stirring for 10 hours. After the starting materialdisappeared as detected by TLC, 10 mL saturated aqueous solution ofNH₄Cl was added, followed by extraction with ethyl acetate twice (in anamount of about 3 mL each time). The organic phases were combined, driedwith anhydrous sodium sulfate, filtered, concentrated, andrecrystallized with ethyl acetate to give Compound 9.

The relevant test data were as follows: ¹H-NMR (300 MHz, CDCl₃): δ=7.20(s, 1H), 7.11 (d, 2H), 6.75 (d, 2H), 6.06 (s, 2H), 3.76 (s, 3H), 2.98(s, 6H); ¹³C-NMR (75 MHz, CDCl₃): δ=170.34, 149.67, 147.66, 142.87,141.34, 131.21, 130.54, 125.76, 124.21, 112.15, 105.44, 101.90, 60.01,40.47.

EXAMPLE 7

Compound 5 (0.01 mmol) obtained in Example 2 was put in a 10 mLround-bottomed flask, followed by addition of 5 mL ethyl ether, andaddition of 0.02 mL 36% aqueous solution of HCl dropwise. The reactionwas carried out at room temperature with stirring for 1 hours. Thereaction solution was filtered. The resulting filter cake was washedwith water for 3 times in an amount of 1 mL each time, and then rapidlywashed with ethyl ether for 3 times in an amount of 1 mL each time.After drying, Compound 10 was obtained.

The relevant test data were as follows: ¹H-NMR (300 MHz, MeOD): δ=7.66(d, 2H), 7.37 (d, 2H), 7.13 (s, 1H), 6.12 (s, 2H), 3.81 (s, 3H), 3.57(s, 3H), 3.36 (s, 6H); ¹³C-NMR (75 MHz, MeOD): δ=168.49, 150.54, 142.97,142.36, 141.70, 140.60, 132.98, 130.34, 126.04, 120.58, 105.59, 103.83,60.43, 52.42, 47.16.

EXAMPLE 8

Compound 5 (0.01 mmol) obtained in Example 2 was put in a 10 mLround-bottomed flask, followed by addition of 5 mL ethyl ether, andaddition of 0.02 mL 40% aqueous solution of HBr dropwise. The reactionwas carried out at room temperature with stirring for 1 hours. Thereaction solution was filtered. The resulting filter cake was washedwith water for 3 times in an amount of 1 mL each time, and then rapidlywashed with ethyl ether for 3 times in an amount of 1 mL each time.After drying, Compound 11 was obtained.

The relevant test data were as follows: ¹H-NMR (300 MHz, CD₃COCD₃):δ=8.01 (d, 2H), 7.37 (d, 2H), 7.09 (s, 1H), 6.18 (s, 2H), 3.81 (s, 3H),3.49 (s, 3H), 3.24 (s, 6H); ¹³C-NMR (75 MHz, CD₃COCD₃): δ=167.41,149.91, 143.14, 141.94, 141.17, 139.82, 132.20, 129.87, 126.29, 121.17,105.13, 103.39, 60.29, 52.07, 46.59.

EXAMPLE 9

Compound 5 (0.01 mmol) obtained in Example 2 was put in a 10 mLround-bottomed flask, followed by addition of benzyl bromide (0.015mmol) and 2 mL toluene. The mixture was heated to 80° C. and thereaction was carried out for 12 hours. The reaction solution wasfiltered. The resulting filter cake was washed with ethyl ether for 3times in an amount of 1 mL each time. After drying, Compound 12 wasobtained.

The relevant test data were as follows: ¹H-NMR (300 MHz, MeOD): δ=7.73(d, 2H), 7.39-7.31 (m, 5H), 7.13 (s, 1H), 7.11 (d, 2H), 6.13 (s, 2H),5.08 (s, 2H), 3.82 (s, 3H), 3.72 (s, 6H), 3.65 (s, 3H); ¹³C-NMR (75 MHz,MeOD): δ=168.50, 150.55, 143.12, 142.49, 141.82, 141.41, 133.89, 132.69,131.88, 130.22, 129.98, 129.01, 125.86, 121.88, 105.72, 103.90, 75.03,60.51, 53.91, 52.48.

EXAMPLE 10 HIV-1 Infectious Titration Test of the Compounds 5-12Obtained in the Above Examples

Titration was conducted according to the improved method of Johnson &Byington: a stock solution of HIV-1_(IIIB) was diluted by four folds ina 96-well plate, in 10 gradients with 6 repeated wells for eachgradient, while setting 6 control wells. To each pore C8166 cells 50 μL(4×10⁵/mL) was added, the final volume being 200 μL per pore. Culturewas conducted at 37° C. and 5% CO₂. On the third day, 100 μL freshRPMI-1640 complete culture medium was replenished. On the seventh day,HIV-1 induced Cytopathic Effect (CPE) in every well was observed usingan inverted microscope, determined by whether or not Syncytium wasformed in every pore. 50% Tissue Culture Infection Dose (TCID₅₀) ofvirus was calculated according to Reed & Muench method.

EXAMPLE 11 Cytotoxicity Test of the Compounds 5-12 Obtained in the AboveExamples on C8166 Host Cell

100 μL of 4×10⁵/mL C8166 cell suspension was mixed with differentcandidate compound solutions. Set three repeated wells. Meanwhile, setcompound-free control wells. Culture was conducted at 37° C. and 5% CO₂for three days. Cell toxicity was tested using MTT colorimetry. OD valuewas tested using EL_(X)800 ELISA instrument, with the measurementwavelength of 595 nm and the reference wavelength of 630 nm. CC₅₀ value(50% Cytotoxic Concentration), i.e. the concentration of compound thatexhibits toxicity to 50% normal T lymphocyte cell line C8166, wascalculated.

EXAMPLE 12 Inhibition Test of the Compounds 5-12 Obtained in the AboveExamples on HIV-1_(IIIB) Induced Cytopathic Effect (CPE) of C8166

8×10⁵/mL C8166 cell suspension was inoculated in 50 μL/well into a96-well cell culture plate containing compound diluted in multipleproportions in 100 μL/well, to which was then added 50 μL ofHIV-1_(IIIB) diluted supernatant (M.O.I. 0.0016). Set three repeatedwells, at the same time, set normal cell control wells that are free ofthe compound. Culture was conducted at 37° C. and 5% CO₂ for three days.Syncytium formation was counted using an inverted microscope (100×).EC₅₀ (50% Effective Concentration), i.e., the concentration of thecompound that inhibits 50% syncytium formation, was calculated.

EXAMPLE 13 Protective Effect of the Compounds 5-12 Obtained in the AboveExamples on HIV-Infected Cells

8×10⁵/mL MT₄ cell suspension was inoculated in 50 μL/well into a 96-wellcell culture plate containing compound diluted in multiple proportionsin 100 μL/well, to half of the wells was added 50 μL of HIV-1_(IIIB)diluted supernatant (M.O.I. 0.0016), and to the other half of the wellswas added 50 μL of the medium. Set two repeated wells per concentrationgradient, and at the same time, set compound-free control wells andblank control wells. Culture was conducted at 37° C. and 5% CO₂. On thethird day, 100 μL of fresh culture medium was replenished per well. Onthe fifth or sixth day, the survival rate of cells was tested using MTTcolorimetry. OD value was tested using EL_(x)800 ELISA instrument, withthe measurement wavelength of 595 nm and the reference wavelength of 630nm. The toxicity of the compounds on normal cells and the protectiveeffect of the compounds on HIV-1_(IIIB) infected cells were calculatedusing formula.

EXAMPLE 14

Calculation method and formula: dose-response curve was plottedaccording to the experimental results; according to Reed & Muenchmethod, 50% effective concentration (EC₅₀) to inhibit viruses, 50%Cytotoxic Concentration (CC₅₀) and anti-HIV-1 activity in theTherapeutic Index (TI) were calculated according to the formula:TI=CC₅₀/EC₅₀.

EXAMPLE 15

Anti-HIV-1 activity test results of the compounds 5-12 obtained in theabove Examples (as shown in Table 1):

TABLE 1 Anti-HIV-1 activity test results of the nitrogen-containingbiphenyl compounds of the present invention toxicity (CC₅₀) Syncytiainhibition Name of (μg/ml) (EC₅₀) (μg/ml) Therapeutic the sample 1 2 1 2index (TI) Compound 5 >1250 411.41 0.44 0.14 >2840.91 Compound 6 >1250358.2 0.49 0.14 >2551.02 Compound 7 880.86 316.56 0.64 0.23 1376.35Compound 8 802.14 245.21 0.93 0.28 862.52 Compound 9 649.14 204.55 >257.88 <25.97 Compound 10 >1250 456.38 0.42 0.15 >2976.19 Compound 1171.94 29.43 0.63 0.26 114.19 Compound 12 53.27 26.59 7.50 3.74 7.10

As can be seen from Table 1, Compound 5 had a very significantanti-HIV-1 activity. Meanwhile, as the substituent group on nitrogenatom of the compound increased, its activity somewhat declined. When theester group of the compound was hydrolyzed into carboxy group (Compound9), its activity declined notably. This indicated that the ester groupimposed a notable impact on the anti-HIV-1 activity of the compound.When Compound 5 was prepared into a hydrochloride salt (Compound 10),the activity was increased, whereas the activity of its ammonium salt orquaternary ammonium salt was declined notably.

EXAMPLE 16

Tablets: one of compounds 5-12 obtained in the above Examples as anactive ingredient 10 mg, lactose 180 mg, starch 55 mg, magnesiumstearate 5 mg.

Preparation: the active ingredient, lactose and starch were mixed, andwetted uniformly with water; the wetted mixture was sieved and dried,and then sieved; magnesium stearate was added, and then the resultingmixture was tabletted. The tablets each weighed 250 mg, and containedthe active ingredient in an amount of 10 mg.

EXAMPLE 17

Ampoules: one of compounds 5-12 obtained in the above Examples as anactive ingredient 2 mg, sodium chloride 10 mg.

Preparation: the active ingredient and sodium chloride were dissolved inan appropriate amount of water for injection, then the resultingsolution was filtered, and filled into ampoules under sterileconditions.

EXAMPLE 18

Capsules: one of compounds 5-12 obtained in the above Examples as anactive ingredient 10 mg, lactose 187 mg, magnesium stearate 3 mg.

Preparation: the active ingredient was mixed with adjuvants, sieved, andmixed uniformly; the resulting mixture was filled into hard gelatincapsules. The capsules each weighted 200 mg, and contained the activeingredient in an amount of 10 mg.

The invention claimed is:
 1. The nitrogen-containing biphenyl compoundhaving the following structural formula (I) or (II):

wherein R₁ is selected from the group consisting of OH, OCH₂Ph and OR;R₂ and R₃ together form —OCH₂O—; R₄ is selected from the groupconsisting of H, OH, OCH₂Ph, OR, and I; R₅ is selected from the groupconsisting of OCH₂Ph and OR; R₆ and R₇ are selected from the groupconsisting of aliphatic hydrocarbyl containing 1-10 carbon atoms; R₈ isselected from the group consisting of H, CH₂Ph and aliphatichydrocarbyl; R₉ is selected from the group consisting of H and aliphatichydrocarbyl; R is selected from the group consisting of aliphatichydrocarbyl and aryl; X is Cl.
 2. The nitrogen-containing biphenylcompound, hich is selected from compounds 5-12 having the followingstructural formulae:


3. A pharmaceutical composition, comprising the nitrogen-containingbiphenyl compound according to claim 1 and at least one pharmaceuticallyacceptable carrier.
 4. A method for preparing the nitrogen-containingbiphenyl compound having the following structural formula (I) or (II):

wherein R₁ is selected from the group consisting of OH, OCH₂Ph and OR;R₂ and R₃ together form —OCH₂O—; R₄ is selected from the groupconsisting of H, OH, OCH₂Ph, OR, and I; R₅ is selected from the groupconsisting of OCH2Ph and OR; R₆ and R₇ are selected from the groupconsisting of aliphatic hydrocarbyl containing 1-10 carbon atoms; R₈ isselected from the group consisting of H, CH₂Ph and aliphatichydrocarbyl; R₉ is selected from the group consisting of H and aliphatichydrocarbyl; R is selected from the group consisting of aliphatichydrocarbyl and aryl; X is Cl, which preparation comprises: couplingCompound 1 and Compound 2 by metallic catalysis to obtain thenitrogen-containing biphenyl compound of the structural formula (I)according to the following reaction formula:

wherein: X′ is F, Cl, Br or I; R′ is an aliphatic or aromatichydrocarbyl, or two R's together form —OC(CH₃)₂—C(CH₃)₂O—; the metalcatalyst is palladium, platinum, rhodium, ruthenium, lithium, copper,magnesium, nickel or zinc; and optionally, the obtainednitrogen-containing biphenyl compound of the structural formula (I) issubjected to a corresponding salt-forming reaction, to obtain thenitrogen-containing biphenyl compound of the structural formula (II). 5.The method for preparing the nitrogen-containing biphenyl compoundaccording to claim 4, comprising mixing Compound 1 and Compound 2,adding Pd(OAc)₂, Bu₄NBr, K₂CO₃ and THF-H₂O (V_(THF)/V_(H2O)=1) to theresultant mixture, heating the mixture to 70° C., reacting for 4 hours,cooling the mixture to room temperature, filtering and concentrating thereaction solution, and purifying by a silica gel column, to obtain thecompound of the structural formula (I).
 6. A method for the treatment ofAIDS, which comprises the step of administering to a patient thenitrogen-containing biphenyl compound according to claim 1.